Differentiation During Encystment and Excystment in Oxytricha fallax*

During encystment of Oxytricha fallax, a wall composed of 4 distinct layers, each derived from a different kind of endoplasmic vesicle, is formed between the 2 unit membranes that cover the vegetative cell. Numerous autophagic vacuoles arise in the endoplasm and later (during excystment undergo internal changes comparable to those characteristic of food vacuoles. Mitochondria aggregate into a band. The 2 macronuclei fuse and their nucleoli become homogeneous. Except for the 2 cell membranes, all visible cortical structures, including cilia, kinetosomes, and microtubules, disappear. Despite the absence of visible ciliature in the mature cyst, the various primordia of the normal vegetative ciliature arise during excystment in the same positional relations to one another as is characteristic of developments during cell division.     

The differentiation of cellular structure during encystment in the soil hypotrichous ciliate Australocirrus cf. australis (Protista,Ciliophora)

Ciliates are able to form resting cysts as a survival strategy in response to stressful environmental factors. Studies on the characteristics of cellular structure during encystment may provide useful information for further understanding of the regulatory mechanism of cellular patterns and supply new clues regarding the phylogeny of ciliates. Scanning and transmission electron microscopies were used to observe the ultrastructure of cells during encystment of the soil ciliate Australocirrus cf. australis. The dedifferentiation of ciliature was revealed for the first time. Ciliary shafts first shortened, and the remaining ciliature, including basal bodies and the fibrillar cirral basket, retracted into the cytoplasm and was surrounded by the autophagic vacuoles and then gradually digested. A large number of autophagic vacuoles were observed in mature resting cysts. Autophagy might not only be necessary for the differentiation of cellular structures during encystment but might also be important to sustain the basic life activities in the resting stage. Australocirrus cf. australis formed a kinetosome-resorbing cyst and contained four layers in the cyst wall: the ectocyst, mesocyst, endocyst and granular layer. The ciliature resorbing state and the number of layers in the cyst wall were consistent with those found in other oxytrichous ciliates. However, the phenomenon wherein the two macronuclear nodules are not fused during encystment is not commonly observed among oxytrichids. Additionally, the octahedral granules in the mesocyst of this species exhibit different morphology from the congeners.     

Mirror-imaged doublets of Tetmemena pustulata: implications for the development of left-right asymmetry

Ciliated protozoa possess cellular axes reflected in the arrangement of their ciliature. Upon transverse fission, daughter cells develop an identical ciliary pattern, ensuring perpetuation of the cellular phenotype. Experimentally manipulated cells can be induced to form atypical phenotypes, capable of intraclonal propagation and regeneration after encystment. One such phenotype in the ciliate Tetmemena pustulata (formerly Stylonychia pustulata) is the mirror-imaged doublet. These cells possess two distinct sets of ciliature, juxtaposed on the surfaces in mirror image symmetry, with a common anterior-posterior axis. We have examined whether individual ciliary components of Tetmemena mirror-image doublets are mirror imaged. Ultrastructural analysis indicates that despite global mirror imaging of the ciliature, detailed organization of the membranelles is reversed in the mirror-image oral apparatus (OA), such that the ciliary effective stroke propels food away from the OA. Assembly of compound ciliary structures of both OAs starts out identically, but as the structures associated with the mirror-image OA continue to form, the new set of membranelles undergoes a 180° planar rotation on the ventral surface relative to the same structures in the typical OA. The overall symmetry of the OA thus appears to be separable from the more localized assembly of individual basal bodies. True mirror imagery of the membranelles would require new enantiomorphic forms of the individual ciliary components, particularly the basal bodies, which is never observed. These observations suggest a mechanistic hypothesis with implications for the development of left-right asymmetry not only in ciliates, but perhaps also in development of left-right asymmetry in general.     

Time Course and Polarity of Primary Phloem Fibre Differentiation in Pisum sativum

Primary phloem fibres of Pisum sativum deposit lignified cellwalls, 2 and 5 days after germination in root and stem, respectively.Fibre bundles reach their final size within 4–6 days.The differentiation of the bundles as a whole is faster in thestem compared with the root. In the special diverging bundlesof the lower internodes of the stem the peripheral portionsmature earlier than the inner portions. A wound in the rootas well as in the stem, interrrupts the differentiation of fibresdirectly below it. At least one bundle below a wound is disturbedand often a whole bundle is missing. The meaning of these findingsconcerning the control of cell size is discussed. Pisum sativum, differentiation, induction, phloem fibres, polarity, time course     

Regulation of Ciliary Pattern in Ciliates*

SYNOPSIS. The modes of pattern regulation found in the ciliates Dileptus and Paraurostyla are compared. These forms are systematically distant but both possess very extensive regenerative capacities. They are characterized by 2 types of ciliary patterns: the ciliature of Dileptus has largely a simple pattern composed of single kinetosomes while that of Paraurostyla has a complex pattern composed of aggregates of kinetosomes interconnected by amorphic matter. In both ciliates a change in cell size evokes pattern regulation which differs substantially in the extent of pattern replacement, timing, and localization of morphogenetic activities. It is concluded that these differences result from the pattern constituents.     

Cyst formation: an important mechanism for the termination of Scrippsiella trochoidea (Dinophyceae) bloom

A sediment trap study was conducted at Daya Bay, South ChinaSea, to investigate the relationships between encystment andpopulation dynamics of Scrippsiella trochoidea from December1999 to January 2001. A dense bloom of S. trochoidea occurredduring the study period from August to September 2000, withthe maximum cell number of 3.18 x 10⁴ cells mL^–1.Two morphotypes of cysts, one with a thick calcareous wall (calcifiedcyst) and another without the obvious calcareous cover (non-calcifiedcyst), were observed during this investigation. The morphologicaland excystment characteristics of these two cyst types werestudied as well. Mass encystments of S. trochoidea, with themaximum of 3.05 x 10⁵ cysts m^–2 d^–1for calcified cyst, and 1.54 x 10⁷ cysts m^–2 d^–1for non-calcified cyst, coincided with the maximum abundanceof the vegetative cells. Encystment caused the transfer of atotal of 2.24–4.49 x 10⁸ cells m^–2 vegetativecells from the water column to the sea bottom during the bloomand resulted in a considerable loss of the bloom population.High assemblages of cysts of S. trochoidea were detected inthe surface sediments as well. This rich ‘seed bed’in the surface sediments caused by the high efficiency of encystmentafter blooms acting as a benthic reservoir for future vegetativepopulation, together with the short dormant period (15–26days) and high germination rate (50–90%), may explainthe repeated occurrence of S. trochoidea blooms in Daya Bay.     

The distribution of dinoflagellate cysts along the Galician (NW Spain) coast

We have studied the distribution of dinoflagellate cysts along10 Galician rías and part of their adjacent continentalshelf. Cyst abundance in the area averaged 856 cysts ml^–1,which is of the same order of magnitude as those found in otherareas of the western European coast. It was higher in the ríasthan in the shelf, having a very heterogeneous distribution,especially in the former. Cyst assemblages in these two areaswere different, suggesting that differences are due to cystproduction rather than to accumulation. Principal componentanalysis, cluster analysis, distribution of macroscopic characteristicsof cyst populations and distribution of single species suggestthat local factors control the distribution in the rías.Nevertheless, a general pattern that splits the whole area intotwo-to the north and to the south of the ría de Camariñas-canbe distinguished. This latter trend was also observed in theshelf and, in our opinion, It should be attributed to threeconcurrent causes: the effect of different upwelling intensitiesor frequencies, the effect of the different numbers and sizesof the ría in each area, and the effect of the presenceof different water masses in these areas. The cyst distributionof a number of individual species was examined and showed threegeneral groups: species with very restricted distribution, suchas Alexandrium sp2 or Scrippsiella sp4, species with a widespreaddistribution along the rías, such as several Scrippsiellaspecies, and species mainly distributed along the shelf, suchas Gymnodinium catenatum. The distribution of cysts belongingto red tide organisms fits quite well with that of their correspondingmotile phases dunng the three previous years for most of theorganisms studied and also during the 1992–1993 period(7–8 years later), but the role of this resting stagein initiating such blooms seems to be highly variable with species.     

Morphology and ontogeny of Tetmemena pustulata indica nov. subspec. (Ciliophora,Hypotricha), from the Thane Creek,Mumbai, India

The morphology and ontogenesis of a novel stylonychid ciliate, Tetmemena pustulata indica nov. subspec., were investigated using live observation and protargol preparation. The new subspecies was isolated from a water sample collected from the Thane Creek, Mumbai, India. At first glance, T. pustulata indica looked very similar in morphology and ontogenesis to the well-known Tetmemena pustulata, however, on detailed investigation some non-overlapping features in the ciliature were identified, i.e., the numbers of cirri in marginal rows, adoral membranelles, and bristles in dorsal kineties. These morphometric differences justified the separation at subspecies level. Furthermore, the resting cysts are different, i.e., with smooth external layer in T. pustulata indica whereas spinous in the nominotypical subspecies T. pustulata pustulata. The Venezuelan population of T. pustulata described by Foissner corresponds very well with the Indian population in the ciliature; however, the information on the resting cyst is lacking for the former. Our study presents an example of a cyst subspecies among hypotrich ciliates, and thus extends the concept which has been mainly reported for spathidiids.     

包囊游仆虫休眠包囊的超微结构研究

包囊游仆虫休眠包囊中,各类纤毛器的纤毛基体上方的大部分纤毛杆退化,或仅保留毛基体,有时部分额腹棘毛的毛基体也瓦解消失。残留纤毛的纤毛杆周围微管和中央微管仍具有“9 2”结构特征,也有少数纤毛杆出现2套“9 2”微管共处于一层纤毛膜内的现象。毛基体中周围三联体微管的中央形成微管形结构聚合体,基体附属结构仅存在基体间连接及纤毛器托架的残余物;非纤毛区皮层表膜下未见微管层。纤毛区皮层含纤毛器腔周围微管层(相当于表膜下微管层)、纤毛器深部及附近的微管束和分散的微管群。并且,纤毛区皮层囊泡内含有呈不同形态的纤毛杆结构;大核核孔明显变大,核孔数目减少,核孔内膜附着染色质。     

The assembly of the FtsZ ring at the mid-chloroplast division site depends on a balance between the activities of AtMinE1 and ARC11/AtMinD1

Chloroplast division comprises a sequence of events that facilitatesymmetric binary fission and that involve prokaryotic-like stromaldivision factors such as tubulin-like GTPase FtsZ and the divisionsite regulator MinD. In Arabidopsis, a nuclear-encoded prokaryoticMinE homolog, AtMinE1, has been characterized in terms of itseffects on a dividing or terminal chloroplast state in a limitedseries of leaf tissues. However, the relationship between AtMinE1expression and chloroplast phenotype remains to be fully elucidated.Here, we demonstrate that a T-DNA insertion mutation in AtMinE1results in a severe inhibition of chloroplast division, producingmotile dots and short filaments of FtsZ. In AtMinE1 sense (overexpressor)plants, dividing chloroplasts possess either single or multipleFtsZ rings located at random intervals and showing constrictiondepth, mainly along the chloroplast polarity axis. The AtMinE1sense plants displayed equivalent chloroplast phenotypes toarc11, a loss-of-function mutant of AtMinD1 which forms replicatingmini-chloroplasts. Furthermore, a certain population of FtsZrings formed within developing chloroplasts failed to initiateor progress the membrane constriction of chloroplasts and consequentiallyto complete chloroplast fission in both AtMinE1 sense and arc11/atminD1plants. Our present data thus demonstrate that the chloroplastdivision site placement involves a balance between the opposingactivities of AtMinE1 and AtMinD1, which acts to prevent FtsZring formation anywhere outside of the mid-chloroplast. In addition,the imbalance caused by an AtMinE1 dominance causes multiple,non-synchronous division events at the single chloroplast level,as well as division arrest, which becomes apparent as the chloroplastsmature, in spite of the presence of FtsZ rings.     

Ultrastructural Observations on Proliferating Storage Cells of Mature Cotyledons of Phaseolus vulgaris L. Cultured in vitro

Explants of mature cotyledons of Phaseolus vulgaris form callusrapidly when cultured in vitro with their adaxial surfaces embeddedin a solidified nutrient medium containing coconut milk, kinetinand 2,4-D. Proliferation is confined to the highly polyploidstorage cells and commences near the adaxial epidermis, whichis soon ruptured by the callus developing internally. Callusformation progresses to the abaxial tissue and within 3–4weeks sub-culturing is possible. The in vitro grown storage cells undergo thinning of their walls,loss of food reserves, hypertrophy, development of various new-wallsand nuclear activation leading to division. The induction ofnuclear and cell divisions within this mature storage tissuecontrasts with normal germination in which these cells undergorapid senescence after depletion of their food reserves. Nuclear division in early callus growth is apparently mainlyamitotic. It is preceded by the development of multiple nucleoli.The nuclear envelope also becomes more complex and deeply lobed;leading to formation of a nuclear isthmus and final separationinto two nuclei. No chromosomes are visible during nuclear fragmentation.Amitosis is accompanied by freely-forming walls, which may developadjacent to a nuclear isthmus and perhaps participate in nuclearfragmentation. Large labyrinthine wall bodies frequently occuron these walls. Mitoses are only observed in already dividedstorage cells. A cell plate forms between the two daughter nuclei,and microtubules are present at its margins in contrast to freely-formingwalls where none are evident. Phaseolus vulgaris L., bean, in-vitro culture, cotyledon, ultrastructure     

Notes on Alexandrium population dynamics

We review within-year and between-year survival strategies ofthe meroplanktonic dinoflagellate Alexandrium, with specialattention to the role of cyst beds and extended dormancy. Someof the constraints on the evolution of cyst bed dynamics arediscussed in the framework of a model borrowed from desert seedecology, in which Q, the annual germination rate, is selectedby p, the probability that the vegetative phase will be successfulon decadal time scales. Since Alexandrium, and the closely relatedPyrodinium, undergo gametogenesis at relatively low cell concentrations,specialized traits must have evolved to achieve syngamy. Itis suggested that motility and the use of chemical signals promotemating, and that the toxins act as pheromones. It is also proposedthat toxins in cysts are used as signals to influence planozygotesettlement so as to control dispersal of this stage, and ensurethat cyst beds are sufficiently stocked to inoculate the watercolumn adequately at the appro priate time of year.     

The Suspensor Haustoria of Some Species of Crotalaria Linn

The structure and function of the suspensor has been studiedin eight species of Crotalaria, in all of which it shows a differentiationinto an upper and a lower part. The peripheral cells of thelower part project out and act as haustorial structures forthe absorption of nutrition from the surrounding endosperm.In G. retusa and G.striata these processes are very long andtubular attacking the surrounding endosperm and nucellar tissuesand extending even into the integumental layers. Remains ofthe suspensor are recognizable even in the mature seed. Thecells of the suspensor possess chloroplasts so that this organalso has a photosynthetic function in addition to its haustorialactivity.     

Histology of Somatic Embryogenesis from Leaf Explants of the Oil Palm Elaeis guineensis

Histological analysis was carried out during the sequence ofevents which lead to the obtaining of somatic embryos of oilpalm. Calluses from the division of perivascular cells formedat the veins of young leaf explants. Subsequent proliferationof histologically similar nodules was by means of a cambium-likezone. Under certain conditions these calluses consisted almostentirely of meristematic cells. They then differentiated rapidly:the cambium-like zone fragmented, leading to protuberance inwhich the cells divide rapidly; epidermal structures were formed,with a network of procambial strands, and synthesis of storagelipids accompanied the formation of these embryo-like structureswhich developed into clumps of true somatic embryos, each witha shoot apex and a root apex. Other structures frequently observedduring in vitro culture are also described and show that alternatepathways do exist. The structure and evolution of somatic embryosare compared to those of zygotic embryos. Storage lipids emergeas an early tracer of the satisfactory development of tissuetowards somatic embryogenesis. Oil palm, Elaeis guineensis, histology, somatic embryogenesis, callogenesis, storage lipids     

Junction Complexes between Sieve Tubes in the Secondary Phloem of Myrtaceae

Junction complexes of unusual structure form between neighbouringsieve tubes in the secondary phloem of Eucalyptus species. Thick-walledribs support thin-walled ‘sieve areas’. In longitudinalsections the structures have a ‘concertina’- likeappearance. They are relatively large, up to 0.2 mm in length.Electron micrographs confirmed that the structures consistedof thin-walled areas perforated with pores, supported by muchthicker ribs. The structures provide a vast surface area fortransfer of metabolites between sieve tubes compared with thatof lateral wall sieve areas of other plants. Hydrolysis of parenchymacell walls occurs during the development of the junction complexes.The structures are only found when sieve tubes are in closeproximity and it is the redifferentiation and partitioning ofintervening parenchyma cells which result in junction complexformation. A survey for the presence of the structures in thephloem of other genera in the family Myrtaceae was made andthey were found in Tristania and Angophora but were not observedin Acmena and Metrosideros. Eucalyptus, sieve tubes, lateral walls, ultrastructure     

Seasonal change of dinoflagellates cyst flux collected in a sediment trap in Omura Bay, West Japan

Sediment trap samples were harvested bimonthly from 1998 to2000 and examined to better understand the species compositionand seasonal variation of dinoflagellate cyst flux in OmuraBay in Japan. Samples’ living cyst flux clearly showedseasonal variation with the higher flux number between autumnand winter. In total, 43 different cyst taxa were recorded,and these were composed of two different ecological groups.The first group included Protoperidinium compressum and Protoperidiniumsubinerme, which increased every autumn to winter. The secondgroup included Gonyaulax spp. and Pheopolykrikos hartmanniiand was trapped throughout the year. These two groups manifestedthe different flux patterns and were, respectively, heterotrophicand autotrophic in nutrition. In the heterotrophic group, protoperidinioidcysts were dominant. Vegetative cells of protoperidinioid areknown to feed mainly diatoms. Sample diatom flux also increasedfrom autumn to winter. Therefore, the increase of protoperidinioidcysts in autumn to winter was observed to correlate with diatomblooms. In contrast, the autotrophic group mostly consistedof Gonyaulacoid cysts and were generally observed throughoutthe year, although occurrence varied between species most likelyresponding to favorable environmental conditions. The resultsindicate that cyst production is closely related to differentnutritional modes.     

Etude Ultrastructurale du CiliéKuklikophrya dragescoi gen. n., sp. n.*

SYNOPSIS The cortical infraciliature of Kuklikophrya dragescoi gen. n., sp. n. is composed of double kinetosomes. Each kinetosome has transverse fibers. The anterior transverse fibers are associated with a sheet of dense material and the posterior transverse fibers are directed toward the posterior part of the body. The posterior kinetosome of a pair has only a short protuberance in the position of the kinetosomal fiber. The cortex has a well developed alveolar layer and a thick ecto-endoplasmic boundary. A distinctive characteristic of the buccal ciliature is the circumoral ciliature whose infraciliature is made up of pairs of cilia-bearing kinetosomes. The antero-posterior polarity of the paroral segment is in inverse relationship to that of the remaining ciliature of the organism. The adoral and preoral ciliary organelles consist of 2 rows of kinetosomes, each of which bears postciliary fibers. A frame of nematodesmata surrounds the cytopharynx which is supported by microtubular bands which impart to it a very specific laminated appearance. The “phagoplasm” is formed by “vermicelli”-like vesicles. The micronucleus is found in the perinuclear area of the macronucleus.     

Inversion of both gating polarity and CO2 sensitivity of voltage gating with D3N mutation of Cx50

The effect of CO₂-induced acidification on transjunctional voltage (V_j) gating was studied by dual voltage-clamp in oocytes expressing mouse connexin 50 (Cx50) or a Cx50 mutant (Cx50-D3N), in which the third residue, aspartate (D), was mutated to asparagine (N). This mutation inverted the gating polarity of Cx50 from positive to negative. CO₂ application greatly decreased the V_j sensitivity of Cx50 channels, and increased that of Cx50-D3N channels. CO₂ also affected the kinetics of V_j dependent inactivation of junctional current (I_j), decreasing the gating speed of Cx50 channels and increasing that of Cx50-D3N channels. In addition, the D3N mutation increased the CO₂ sensitivity of chemical gating such that even CO₂ concentrations as low as 2.5% significantly lowered junctional conductance (G_j). With Cx50 channels G_j dropped by 78% with a drop in intracellular pH (pH_i) to 6.83, whereas with Cx50-D3N channels G_j dropped by 95% with a drop in pH_i to just 7.19. We have previously hypothesized that the way in which V_j gating reacts to CO₂ might be related to connexin’s gating polarity. This hypothesis is confirmed here by evidence that the D3N mutation inverts the gating polarity as well as the effect of CO₂ on V_j gating sensitivity and speed. cell communication; lens; gap junctions; chemical gating; channel gating; Xenopus oocytes     

The Fine-Structural Localization of Phosphatases in Neurosecretory Cells Within the Ganglia of Certain Gastropod Snails

The neurosecretory cells in the cerebral ganglia of the snail,Planorbis trivolvis, have been examined for structural detailsand distribution of phosphatase activities. A more preliminarystudy on the localization of phosphatases in the neurosecretorycells of Helix aspersa has also been made. These neurons inboth species contain typical elementary neurosecretory granuleswhich appear to be elaborated or condensed in the Golgi saccules.Immature elementary granules are identifiable as "primary lysosomes"in that they contain acid phosphatase activity as well as nucleosidephosphatases. The mature elementary granules display no phosphataseactivity. Some saccules of the Golgi are also reactive for severalnucleoside phosphatases, including thiamine pyrophosphatase(TPPase). In Planorbis, TPPase is also present in the cisternaeof the endoplasmic reticulum. Larger, membrane bound, electron-densebodies (lipochondria) are found in close spatial associationwith the Golgi region. These also possess acid phosphatase;in addition, they contain nucleoside diphosphatases and (inPlanorbis) adenosine triphosphatase. Their content of acid phosphataseand the features of their fine structure indicate that theyare lysosomes akin to the dense bodies of vertebrate neurones. The significance and implications of these results are considered,as are other details of the neuronal and glial structure inPlanorbis.